产品介绍 | | |
Mouse anti Human CD152 antibody, clone BNI3recognizes human CD152, also known as CTLA-4 (cytotoxic T-lymphocyte-associated antigen 4), an inhibitory receptor and negative regulator of T-cell responses. CD152 is a single pass type 1 transmembrane protein belonging to the immunoglobulin superfamily containing a singleIg-v-likedomain in the extracellular region.CD152 along with CD28 binds to the co-stimulatory molecules CD80 and CD86 (Azumaet al.1993).Mouse anti human CD152 antibody, clone BNI3 is able to block ligand binding on the Raji B-cell line (Steineret al.2001) and blocks binding of an alternative clone, BNI8 to CTLA-4/Ig in ELISA. Mouse anti Human CD152 antibody, clone BNI3 binds to the same epitope as classified anti CTLA-4 clones 11D4 and 10A8 (Wanget al.In: Leukocyte typing VI 1997 Garland Publishing Inc. pp97-98,Bull World Health Organ. 1997).The cytoplasmic domain of CD152 contains a critical tyrosine at residue 201 phosphorylated by Janus Kinase 2 which subsequently controls surface expression through regulation of CD152 interaction with AP-2 (Shiratoriet al.1997,Chikumaet al.2000). CD152 is expressed primarily as an intracellular antigen with transport to the cell surface under tight regulation of several molecules including Trim, PLD and TIRC7, CD152 also demonstrates rapid internalization once expressed at the cell surface (Valket al.2008). CD152 plays a significant role in maintaining tolerance to self antigens and defects in CD152 presentation and expression has been implicated in a number of autoimmune diseases (Romo-Tenaet al.2013).This antibody can be used to develop a pharmacokinetic (PK) bridging assay to measure free fusion protein drug abatacept. It is recommended as the capture reagent, paired with human anti abatacept antibodyHCA335as the detection antibody.Abatacept (Orencia) is a recombinant fusion protein consisting of the extra-cellular domain of human CTLA-4, fused to the modified Fc (hinge, CH2, and CH3 domains) portion of human IgG1. |
产品详情 | | |
Target Species | Human |
Product Form | Purified IgG - liquid |
Preparation | Purified by affinity chromatography on Protein A from tissue culture supernatant |
Buffer Solution | Phosphate buffered saline |
Preservative Stabilisers | <0.1% Sodium Azide (NaN3) |
Immunogen | Human CTLA-4/human IgG heavy chain fusion protein. |
Approx. Protein Concentrations | IgG concentration 1.0 mg/ml |
Fusion Partners | Spleen cells from immunised BALB/c mice were fused with cells of the mouse P3/X63-Ag8.653 myeloma cell line. |
Regulatory | For research purposes only |
Guarantee | Guaranteed for 12 months from the date of despatch or until the date of expiry, whichever comes first. Please see label for expiry date. |
Acknowledgements | Orencia is a trademark of Bristol-Myers Squibb Company |
存储条件 | | |
Store at +4°C. DO NOT FREEZE.This product should be stored undiluted. Should this product contain a precipitate we recommend microcentrifugation before use. |
应用 | | |
Application Name | Verified | Min Dilution | Max Dilution |
ELISA | √ | | |
Flow Cytometry1 | √ | Neat | 1/10 |
Immunofluorescence | √ | | |
Immunohistology - Frozen2 | √ | 5ug/ml | 50ug/ml |
Immunohistology - Paraffin | X | | |
Immunoprecipitation | √ | | |
This product has been reported to work in the following applications. This information is derived from testing within our laboratories, peer-reviewed publications or personal communications from the originators. Please refer to references indicated for further information. For general protocol recommendations, please visit theantibody protocolspage.1Membrane permeabilization is required for this application. Bio-Rad recommend the use of Leucoperm™(Product CodeBUF09) for this purpose.2The epitope recognised by this antibody is reported to be sensitive to formaldehyde fixation and tissue processing. Bio-Rad recommends the use of acetone fixation for frozen sections.Where this antibody has not been tested for use in a particular technique this does not necessarily exclude its use in such procedures. Suggested working dilutions are given as a guide only. It is recommended that the user titrates the antibody for use in their own system using appropriate negative/positive controls.- Flow CytometryUse 10ul of the suggested working dilution to label 1x106cells in 100ul.
- ELISAClone BNI3 can be used as the capture antibody in a bridging ELISA withHCA335as the detection antibody.
|
数据 |
Human Anti-Abatacept Antibody bridging ELISA for Pharmacokinetic (PK) assay developmentA microtiter plate was coated overnight with Mouse Anti-Human CD152 Antibody, clone BNI3 (MCA1724) at a concentration of 1 µg/ml. After washing and blocking with PBST + 5% BSA, 10% human serum was added spiked with increasing concentrations of abatacept. Detection was performed using HRP conjugated Human Anti-Abatacept Antibody, clone AbD37058ia (HCA335) at a concentration of 2 µg/ml in HISPEC Assay Diluent (BUF049A) followed by QuantaBlu Fluorogenic Peroxidase Substrate. Data are shown as the mean of three measurements. HRP conjugation of the antibody used in this assay was performed using a LYNX Rapid HRP Antibody Conjugation Kit (LNK002P). |
同型 |
Description | Product Code | Applications | Pack Size |
Mouse IgG2a Negative Control | MCA929 | Flow | 100 Tests |
相关产品 |
Description | Product Code | Applications | Pack Size |
Human anti Abatacept | HCA335 | ELISA | 0.1 mg |
Human anti Abatacept | HCA336 | ELISA | 0.1 mg |
文献 |
1. Castan, J.et al.(1997) Accumulation of CTLA-4 expressing T lymphocytes in the germinal centres of human lymphoid tissues.Immunology. 90 (2): 265-71.2. Lee, C.C.et al.(2009) The regulatory function of umbilical cord blood CD4(+) CD25(+) T cells stimulated with anti-CD3/anti-CD28 and exogenous interleukin (IL)-2 or IL-15.Pediatr Allergy Immunol. 20 (7): 624-32.3. Pistillo, M.P.et al.(2003) CTLA-4 is not restricted to the lymphoid cell lineage and can function as a target molecule for apoptosis induction of leukemic cells.Blood. 101: 202-9.4. Tan, P.H.et al.(2005) Creation of tolerogenic human dendritic cells via intracellular CTLA4: a novel strategy with potential in clinical immunosuppression.Blood. 106: 2936-43.5. Steiner, K.et al.(2001) Increased expression of CTLA-4 (CD152) by T and B lymphocytes in Wegener's granulomatosis.Clin Exp Immunol. 126: 143-50.6. Lu, C.H.et al.(2016) DNA Methyltransferase Inhibitor Promotes Human CD4+CD25hFOXP3+Regulatory T Lymphocyte Induction under Suboptimal TCR Stimulation.Front Immunol. 7: 488.7. Steiner, K.et al.(1999) Enhanced expression of CTLA-4 (CD152) on CD4+ T cells in HIV infection.Clin Exp Immunol. 115 (3): 451-7.8. Ward, F.J.et al.(2013) The soluble isoform of CTLA-4 as a regulator of T-cell responses.Eur J Immunol. 43 (5): 1274-85. |